The effect of aqueous humor ascorbate on ultraviolet-B-induced DNA damage in lens epithelium.
OBJECTIVE
High levels of ascorbic acid are known to be present in the aqueous humor of many diurnal species, whereas nocturnal animals have low concentrations of the compound. The purpose of this study was to test the hypothesis that the high concentration of aqueous ascorbate in diurnal animals protects the lens against ultraviolet (UV)-induced damage to the eye. This study compares the effect of UV-B-induced DNA strand breaks on the lens epithelia of guinea pigs and rats after depletion or elevation of aqueous humor ascorbate, respectively.


METHODS
Eyes of guinea pigs and rats were exposed to UV-B radiation (0.25-0.75 J/cm2 on the cornea) for 10 minutes, and DNA strand breaks in lens epithelium were measured by single-cell gel electrophoresis. Ascorbic acid concentration in the aqueous humor, lens, and lens-capsule epithelium were assayed by spectrophotometric and electrochemical methods. For depletion of aqueous humor and lens ascorbate in guinea pigs, the animals were maintained on an ascorbate-deficient diet. Aqueous ascorbic acid was elevated in the rat by intraperitoneal injections of sodium ascorbate (1 g/kg).


RESULTS
The ascorbate concentration in the aqueous humor of the normal rat was approximately 3% that of the guinea pig, whereas the concentration of the compound in the lens of the normal rat was 10% that of the guinea pig. Guinea pigs fed an ascorbate-deficient diet showed a dramatic drop of more than 80% in aqueous humor ascorbate in the first week, whereas lens ascorbate decreased by approximately 25% during this time period. After a single intraperitoneal injection of sodium ascorbate in the rat, aqueous humor ascorbic acid increased nearly 30 times that in the control, whereas lens ascorbate increased by approximately 30%. The extent of DNA damage in the lens epithelium of a normal rat exposed to UV-B was significantly greater than that occurring in lenses of normal guinea pigs after exposure to the same dose of radiation. Lenses from ascorbate-deficient guinea pigs showed 50% more DNA damage than those from normal guinea pigs after UV exposure, whereas the lenses in ascorbate-injected rats exhibited significant protection against UV-induced DNA strand breaks.


CONCLUSIONS
High levels of ascorbic acid in the aqueous humor had a protective effect against UV-induced DNA damage to lens epithelium. The results were consistent with the hypothesis that high ascorbic acid in diurnal animals protects the lens against the cataractogenic effect of UV radiation in sunlight.
Reddy
V N
VN
Eye Research Institute, Oakland University, Rochester, Michigan 48309-4401, USA.
Giblin
F J
FJ
Lin
L R
LR
Chakrapani
B
B
D000818:Animals; D001082:Aqueous Humor / Q000737:chemistry / Q000502:physiology*; D001205:Ascorbic Acid / Q000032:analysis / Q000494:pharmacology / Q000502:physiology*; D001206:Ascorbic Acid Deficiency / Q000378:metabolism / Q000503:physiopathology; D004249:DNA Damage / Q000528:radiation effects*; D004032:Diet; D004848:Epithelium / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D006168:Guinea Pigs; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D008297:Male; D011833:Radiation Injuries, Experimental / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014466:Ultraviolet Rays / Q000009:adverse effects*
D001205:Ascorbic Acid